Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Western Blotting

Western Blotting

Western blotting is an analytical technique for protein identification. It has various applications in immunology and medicine, including detecting diseases like bovine spongiform encephalopathy, mad cow disease, and human and feline immunodeficiency virus from biological samples.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.

SDS-PAGE

SDS-PAGE

Gel electrophoresis is a method that separates biological macromolecules like nucleic acids or proteins by forcing them to pass through a gel matrix under an electric field.
A variation of gel electrophoresis, termed polyacrylamide gel electrophoresis (PAGE), is commonly used for separating proteins according to their molecular size by passing them through a polyacrylamide gel. Because of the varying charges associated with amino acid side chains, PAGE can be used to separate intact proteins...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

<i>Proteomes</i> Annual Report Card 2025.

Proteomes·2026

Same author

Cold-stored platelets: revisiting assumptions and addressing variability to support implementation.

Frontiers in medicine·2025

Same author

Peptide ratios for post-mortem interval estimation using targeted liquid chromatography triple quadrupole mass spectrometry.

International journal of legal medicine·2025

Same author

Vitamin C attenuates low-level PM<sub>2.5</sub> exposure-induced lung inflammation and mitochondrial loss.

Environment international·2025

Same author

Sterols are key to coral larvae survival, swimming capacity, and thermal tolerance.

Communications biology·2025

Same author

Matrix directs trophoblast differentiation in a bioprinted organoid model of early placental development.

Nature communications·2025

Search research articles

Related Experiment Video

Updated: May 14, 2026

Fast and Sensitive Colloidal Coomassie G-250 Staining for Proteins in Polyacrylamide Gels

Fast and Sensitive Colloidal Coomassie G-250 Staining for Proteins in Polyacrylamide Gels

Published on: August 3, 2009

Coomassie blue staining for high sensitivity gel-based proteomics.

Victoria J Gauci¹, Matthew P Padula, Jens R Coorssen

¹Molecular Physiology, and the Molecular Medicine Research Group, School of Medicine, University of Western Sydney, Sydney, Australia.

Journal of Proteomics

|February 23, 2013

Summary

Optimized colloidal coomassie blue (CCB) staining offers sensitive, quantitative protein analysis. The Neuhoff formulation (NCCB) provides superior 2D gel spot detection at a lower cost compared to fluorescent stains.

Keywords:

1DE 2DE AS BGAL CA Coomassie Fluorescence Gel electrophoresis IPV IRFD LDR LLD LYS MS NCCB Neuhoff colloidal coomassie blue PHOSB Protein staining S(2)/BG SR STI SYPRO Ruby TYRP Top-down proteomics ammonium sulphate beta-galactosidase carbonic anhydrase infrared fluorescent detection inter-protein variability linear dynamic range lowest limit of detection lysozyme mass spectrometry one dimensional gel electrophoresis phosphorylase b signal squared to background ratio soybean trypsin inhibitor trypsinogen two dimensional gel electrophoresis

More Related Videos

Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid

Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid

Published on: August 14, 2009

Staining Proteins in Gels

Staining Proteins in Gels

Published on: July 8, 2008

Related Experiment Videos

Last Updated: May 14, 2026

Fast and Sensitive Colloidal Coomassie G-250 Staining for Proteins in Polyacrylamide Gels

Fast and Sensitive Colloidal Coomassie G-250 Staining for Proteins in Polyacrylamide Gels

Published on: August 3, 2009

Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid

Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid

Published on: August 14, 2009

Staining Proteins in Gels

Staining Proteins in Gels

Published on: July 8, 2008

Area of Science:

Proteomics
Biochemistry
Analytical Chemistry

Background:

Gel electrophoresis (1DE and 2DE) are standard proteome analysis methods.
Protein detection typically uses stains like colloidal coomassie blue (CCB) or SYPRO Ruby (SR).
Fluorescent stains offer higher sensitivity and dynamic range, but CCB also possesses fluorescent properties.

Purpose of the Study:

To optimize CCB staining protocols for enhanced protein detection sensitivity and quantitative analysis.
To compare the performance of BioSafe and Neuhoff formulations (NCCB) of CCB against SYPRO Ruby (SR).
To evaluate the cost-effectiveness of optimized CCB staining for proteomic analysis.

Main Methods:

Optimization of staining protocols for BioSafe and NCCB colloidal coomassie blue formulations.
Infrared detection of CCB-stained proteins.
Two-dimensional gel electrophoresis (2DE) analysis of mouse brain and Arabidopsis thaliana proteomes.
Comparison of spot detection sensitivity and dynamic range with SYPRO Ruby (SR).

Main Results:

Optimized CCB staining protocols improved selectivity without compromising sensitivity.
The Neuhoff formulation (NCCB) demonstrated superior spot detection in 2D gels compared to SYPRO Ruby (SR).
Linear dynamic range for BioSafe and NCCB was comparable, and better than SR.
NCCB offers a cost-effective alternative for sensitive, quantitative in-gel protein analysis.

Conclusions:

Optimized NCCB staining provides a highly sensitive and quantitative method for 2D gel electrophoresis.
NCCB is a superior and more economical choice for proteomic analysis compared to traditional fluorescent stains like SR.
This optimized staining protocol enables enhanced protein spot detection in complex proteomes.